Analytical Method For Active Ingredients Of Scutellaria Baicalensis Georgi And The Identification Of Its Producing Areas

Scutellaria baicalensis Georgi (SBG) is a kind of traditional Chinese medicine, which has been widely used in traditional Chinese prescriptions, and has been used as a kind of food additive. It shows effective biological activity in many aspects. At present, most of researchs about SBG were focused on the flavonoids in the root of SBG, and the studies about the active substances in the aerial part of SBG were a few. Meanwhile, as reported in relevant papers, the trace amounts of organic acid compounds in SBG showed strong activity, which could be considered as another active ingredients in SBG. In order to roundly investigate the active ingredients in SBG, and accurately assess the quality of herbal, in this thesis, three analytical methods were developed for the determination of two kinds of active ingredients in SBG, and a comprehensive fingerprint library of SBG which were from different localities was established. In addition, the similarity evaluation and cluster analysis were applied to identify the SBG from different habitats, which has great significance for the quality evaluation of SBG.The following work has been mainly finished in this study:1.A fingerprint analytical method was developed for the determination of SBG which were from different habitats by high performance liquid chromatograghy-diode array detector (HPLC-DAD). The contents include:(1)In order to make the HPLC fingerprint analysis method have the characteristics, stability, systematization and reliability, the extract methods for SBG, the chromatographic conditions and the validation of methodology, etc. had been investigated.(2)Fingerprints of the roots, stems and leaves of SBG under optimal conditions were qualitatively analyzed by the combination of chromatographic retention and the data characteristics of UV spectra.(3)The contents of baicalin in the roots of SBG which were from30different habitats and in the stems and leaves of SBG from16different habitats were systematically analyzed.(4)The fingerprints of SBG from different habitats were introduced into the "similarity evaluation system for chromatographic fingerprint of TCM", and the matching of fingerprints was analysed. Finally, it was successfully developed the fingerprints spectrum library of the roots, stems and leaves of SBG from different habitats.2.A high performance liquid chromatograghy/electrospray-mass spectrometry(HPLC/ESI-MS) method was developed for the determination of trace amounts of phenolic acids in the roots, stems and leaves of SBG. The contents include:(1)In order to make the analytical method have characteristics of high sensitivity, high accuracy, good repetitiveness and good stability, the optimization of the extract methods of organic acid in the roots, stems and leaves of SBG, the optimization of HPLC/ESI-MS conditions and the validation of methodology had been investigated.(2)Target ions were analyzed by secondary mass spectrometry, and the information of all peaks achieves the global qualitative analysis according to the characteristics of parent ions, the secondary mass spectrometry fragment ions and chromatographic retention. Finally, six kinds of organic acids had been identified from the roots, stems and leaves of SBG.(3)The organic acids which had higher activity in SBG were quantitatively analyzed, and the method had been successfully applied to the analysis of the authentic ingredients from Chengde, Hebei.3.A new desorption corona beam ionization coupling with ion trap mass spectrometry (DCBI-MS) analytical method was developed for the rapid identification of flavonoids in the roots of SBG. The contents include:(1)The vital factors for influence the mass spectrometry response signal, such as the sample preparation methods, mass spectrometry monitoring mode, and ion source desorption temperature, etc. were inverstigated. And the optimal condition for high-throughput and high sensitivity analysis of DCBI-MS were obtained.(2)Target ions were analyzed by secondary mass spectrometry, and the information of all peaks achieves the global qualitative analysis according to the characteristics of parent ions and the secondary mass spectrometry fragment ions. Finally, twelve kinds of flavonoid aglycones of the roots had been identified.(3)The new DCBI-MS method had been compared with the universal HPLC-UV analysis method, to achieve complementary advantages, and it had been successfully applied in the analysis of SBG which were from eleven different habitats.4.The study was applied to identify the differences of the herbal ingredients which were from different habitats based on HPLC fingerprints. The contents include:(1)The fingerprints of the roots, stems and leaves of SBG which were from different habitats were analyzed, and then similarity matching higher batches(>0.85) were selected to establish the common pattern of three types of samples to compare the differences of fingerprints.(2)Using SPSS statistical software, the HPLC fingerprints about the roots of SBG which were from30different habitats were identified by the cluster analysis.(3)Using SPSS statistical software, HPLC fingerprints of the roots, stems and leaves of SBG from16different habitats were conducted to do the cluster analysis. Finally, the comprehensive fingerprint identification system had been established to identify medicinal authenticity, origin differences and evaluate the herbal quality.